Nanoelectroablation of Murine Tumors Triggers a CD8-Dependent Inhibition of Secondary Tumor Growth.
Nanoelectroablation of Murine Tumors Triggers a CD8-Dependent Inhibition of Secondary Tumor Growth.
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DOI：
10.1371/journal.pone.0134364
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Nuccitelli P
中科院分区：
文献类型：
--
作者：
Nuccitelli R;Berridge JC;Mallon Z;Kreis M;Athos B;Nuccitelli P
我们使用大鼠原位肝细胞癌模型和小鼠同种异体移植肿瘤模型来研究纳秒脉冲电场（nsPEF）的肝肿瘤消融。我们证实，nsPEF治疗触发大鼠肝肿瘤细胞的凋亡，如在治疗后两小时内出现的裂解的半胱天冬酶3和9所示。此外，我们提供的证据表明，nsPEF治疗导致钙网蛋白（CRT）易位到细胞表面，这被认为是一种损伤相关的分子模式，指示免疫原性细胞死亡。我们提供了直接的证据，纳米电消融触发了CD8依赖性抑制继发性肿瘤的生长，通过比较nsPEF治疗的大鼠中的继发性原位肝肿瘤的生长速度与nsPEF治疗的大鼠中的CD8+细胞毒性T细胞耗尽。与CD8耗竭大鼠中的肿瘤生长相比，这些继发性肿瘤的生长受到严重抑制，在相同的一周生长期后，其平均大小仅为原发性肿瘤大小的3%。相比之下，当我们耗尽CD8+ T细胞时，第二个肿瘤生长得更强劲，达到第一个肿瘤大小的54%。此外，我们用免疫组化证明，CD8+ T细胞高度富集在表现出缓慢生长的继发性肿瘤中。我们还表明，用nsPEF处理的等基因肿瘤细胞接种小鼠可刺激免疫应答，以CD8+依赖性方式抑制继发性肿瘤的生长。我们得出结论，纳米电消融触发了CD8+细胞毒性T细胞的产生，导致继发性肿瘤生长的抑制。
We have used both a rat orthotopic hepatocellular carcinoma model and a mouse allograft tumor model to study liver tumor ablation with nanosecond pulsed electric fields (nsPEF). We confirm that nsPEF treatment triggers apoptosis in rat liver tumor cells as indicated by the appearance of cleaved caspase 3 and 9 within two hours after treatment. Furthermore we provide evidence that nsPEF treatment leads to the translocation of calreticulin (CRT) to the cell surface which is considered a damage-associated molecular pattern indicative of immunogenic cell death. We provide direct evidence that nanoelectroablation triggers a CD8-dependent inhibition of secondary tumor growth by comparing the growth rate of secondary orthotopic liver tumors in nsPEF-treated rats with that in nsPEF-treated rats depleted of CD8+ cytotoxic T-cells. The growth of these secondary tumors was severely inhibited as compared to tumor growth in CD8-depleated rats, with their average size only 3% of the primary tumor size after the same one-week growth period. In contrast, when we depleted CD8+ T-cells the second tumor grew more robustly, reaching 54% of the size of the first tumor. In addition, we demonstrate with immunohistochemistry that CD8+ T-cells are highly enriched in the secondary tumors exhibiting slow growth. We also showed that vaccinating mice with nsPEF-treated isogenic tumor cells stimulates an immune response that inhibits the growth of secondary tumors in a CD8+-dependent manner. We conclude that nanoelectroablation triggers the production of CD8+ cytotoxic T-cells resulting in the inhibition of secondary tumor growth.